Treatment of ocular hypertension with a synergistic combination

ABSTRACT

A method for treatment of ocular hypertension which comprises oculary administering, to a subject in need of such treatment, a oculo-hypotensively synergistic combination of 
     (a) a 13,14-dihydro-15-ketoprostaglandin compound, and 
     (b) a β-adrenergic blocker 
     in an amount effective in treatment of ocular hypertension.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a treatment of ocular hypertension witha synergistic combination comprising (a) a13,14-dihydro-15-ketoprostaglandin compound and (b) a β-adrenergicblocker.

The compounds used as the component (a) in the present invention areprostaglandin analogues which can be obtained synthetically.

2. Information of Prior Art

Prostaglandins (hereinafter, prostaglandins are referred to as PGs) aremembers of a class of organic carboxylic acid that are contained inhuman and most other mammalian tissues or organs and that exhibit a widerange of physiological activities. Naturally occurring PGs possess as acommon structural feature the prostanoic acid skeleton: ##STR1## Somesynthetic analogues have somewhat modified skeletons. The primary PGsare classified based on the structural feature of the five-memberedcycle moiety into PGAs, PGBs, PGCs, PGDs, PGEs, PGFs, PGGs, PGHs, PGIsand PGJs, and also on the presence or absence of unsaturation andoxidation in the chain moiety as:

Subscript 1: 13,14-unsaturated-15-OH

Subscript 2: 5,6- and 13,14-diunsaturated-15-OH

Subscript 3: 5,6- 13,14- and 17,18-triunsaturated-15-OH

Further, PGFs are sub-classified according to the configuration ofhydroxy group at position 9 into α(hydroxy group being in the alphaconfiguration) and β(hydroxy group being in the beta configuration).

The fact that the above compounds under item (a) have ocular hypotensiveactivity has been known by Japanese Patent Publication No. A-108/1990.It has also been described in Japanese Patent Publication No.A-313728/1988, page 7, column 3, line 7 from bottom to page 8, column 4,line 4, that a combination of PGF₂ α isopropyl ester and Timolol (anagent for treating glaucoma) may be advantageous because the ocularhypotensive activity of the former is not inhibited by a β-adrenergicblocker such as the latter. Such description, however, neither show acombined use of the β-adrenergic blocker and the component (a) in thepresent invention nor suggest that said combined use may synergisticincrease in effect or decrease in side-effect because PGF₂ α is aprimary PG having a trans double bond between positions 13 and 14, ahydroxy group (in α-conformation) at position 15 and 20 carbon atoms inthe basic structure, while the 13,14-dihydro-15-keto-PGs are compoundshaving a saturated bond between positions 13 and 14, an oxo group atposition 15 in place of a hydroxy group which has been considered as animportant group for activities of primary PGs and thus significantlydifferent from the primary PGF₂ α.

After an extensive study on the possibility that the effect of thecomponent (a) in the present invention is improved by combining it witha variety of compounds, the present inventor has surprisingly discoveredthat the effect of the component (a) is significantly improved andside-effect is decreased by co-administration with a β-adrenergicblocker such as Timolol. Said discovery leads to the present invention.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides a method for treatmentof ocular hypertension which comprises ocularly administering, to asubject in need of such treatment, an oculo-hypotensively synergisticcombination of

(a) a 13,14-dihydro-15-ketoprostaglandin compound, and

(b) a β-adrenergic blocker

in an amount effective in treatment of ocular hypertension.

In a second aspect, the present invention provides a use of anoculo-hypotensively synergistic combination of

(a) a 13,14-dihydro-15-ketoprostaglandin compound, and

(b) a β-adrenergic blocker

for the manufacture of a medicament useful in treatment of ocularhypertension.

In a third aspect, the present invention provides a pharmaceuticalcomposition for treatment of ocular hypertension which comprising anoculo-hypotensively synergistic combination of

(a) a 13,14-dihydro-15-ketoprostaglandin compound, and

(b) a β-adrenergic blocker

in association with a pharmaceutically acceptable carrier, diluent orexcipient.

DETAILED DESCRIPTION OF THE INVENTION

The "13,14-dihydro-15-ketoprostaglandin compounds", used as thecomponent (a) in the present invention and referred to as the component(a), include any prostaglandin derivatives which have a single bond inplace of the double bond between positions 13 and 14 and an oxo group inplace of the hydroxy group at position 15 of the prostanoic acidnucleus.

Nomenclature

Nomenclature of the component (a) herein uses the numbering system ofprostanoic acid represented in formula (A) shown above.

While formula (A) shows a basic skeleton having twenty carbon atoms, the13,14-dihydro-15-keto-PG compounds used in the present invention are notlimited to those having the same number of carbon atoms. The carbonatoms in Formula (A) are numbered 2 to 7 on the α-chain starting fromthe α-carbon atom adjacent to the carboxylic carbon atom which isnumbered 1 and towards the five-membered ring, 8 to 12 on the said ringstarting from the carbon atom on which the α-chain is attached, and 13to 20 on the ω-chain starting from the carbon atom adjacent to the ring.When the number of carbon atoms is decreased in the α-chain, the numberis deleted in order starting from position 2 and when the number ofcarbon atoms is increased in the α-chain, compounds are named assubstituted derivatives having respective substituents at position 1 inplace of carboxy group (C-1). Similarly, when the number of carbon atomsis decreased in the ω-chain, the number is deleted in order startingfrom position 20 and when the number of carbon atoms is increased in theω-chain, compounds are named as substituted derivatives havingrespective substituents at position 20. Stereochemistry of the compoundsis the same as that of above formula (A) unless otherwise specified.Thus, 13,14-dihydro-15-keto-PG compounds having 10 carbon atoms in theω-chain is nominated as 13,14-dihydro-15-keto-20-ethyl-PGs.

The above formula expresses a specific configuration which is the mosttypical one, and in this specification compounds having such aconfiguration are expressed without any specific reference to it.

In general, PGDs, PGEs and PGFs have a hydroxy group on the carbon atomat position 9 and/or 11 but in the present specification the term"13,14-dihydro-15-keto-PG compounds" includes PGs having a group otherthan a hydroxyl group at position 9 and/or 11. Such PGs are referred toas 9-dehydroxy-9-substituted-PG compounds or11-dehydroxy-11-substituted-PG compounds.

As stated above, nomenclature of the component (a) is based upon theprostanoic acid. These compounds, however, can also be named accordingto the IUPAC naming system. For example,13,14-dihydro-15-keto-16R,S-fluoro-PGE₂ is(Z)-7-{(1R,2R,3R)-3-hydroxy-2-[(4R,S)-fluoro-3-oxo-1-octyl]-5-oxocyclopentyl}-hept-5-enoicacid. 13,14-dihydro-15-keto-20-ethyl-PGE₂ is(Z)-7-{(1R,2R,3R)-3-hydroxy-2-[3-oxo-1-decyl]-5-oxocyclopentyl}-hept-5-enoicacid. 13,14-dihydro-15-keto-20-ethyl-PGF₂ α isopropyl ester is isopropyl(Z)-7-[(1R,2R,3R,5S)-3,5-dihydroxy-2-(3-oxo-1-decyl)-cyclopentyl]-hept-5-enoate.13,14-dihydro-15-keto-20-methyl-PGF₂ α methyl ester is methyl(Z)-7-[(1R,2R,3R,5S)-3,5-dihydroxy-2-{3-oxo-1-nonyl}-cyclopentyl]-hept-5-enonate.

Preferred Compounds

The component (a) used in the present invention may be any derivativesof PG insofar as they are saturated between positions 13 and 14 and havean oxo group at position 15 in place of the hydroxy group, and may haveno double bond (PG subscript 1 compounds), a double bond betweenpositions 5 and 6 (PG subscript 2 compounds), or two double bondsbetween positions 5 and 6 as well as positions 17 and 18 (PG subscript 3compounds).

Typical examples of the compounds used in the present invention are13,14-dihydro-15-keto-PGA₁, 13,14-dihydro-15-keto-PGA₂,13,14-dihydro-15-keto-PGA₃, 13,14-dihydro-15-keto-PGB₁,13,14-dihydro-15-keto-PGB₂, 13,14-dihydro-15-keto-PGB₃,13,14-dihydro-15-keto-PGC₁, 13,14-dihydro-15-keto-PGC₂,13,14-dihydro-15-keto-PGC₃, 13,14-dihydro-15-keto-PGD₁,13,14-dihydro-15-keto-PGD₂, 13,14-dihydro-15-keto-PGD₃,13,14-dihydro-15-keto-PGE₁, 13,14-dihydro-15-keto-PGE₂,13,14-dihydro-15-keto-PGE₃, 13,14-dihydro-15-keto-PGF₁,13,14-dihydro-15-keto-PGF₂, 13,14-dihydro-15-keto-PGF₃, wherein PG is asdefined above as well as their substitution products or derivatives.

Examples of substitution products or derivatives includepharmaceutically or physiologically acceptable salts and esters at thecarboxy group at the alpha chain, unsaturated derivatives having adouble bond or a triple bond between positions 2 and 3 or positions 5and 6, respectively, substituted derivatives having substituent(s) oncarbon atom(s) at position 3, 5, 6, 16, 17, 19 and/or 20 and compoundshaving lower alkyl or a hydroxy (lower) alkyl group at position 9 and/or11 in place of the hydroxy group, of the above PGs.

Examples of substituents present in preferred compounds are as follows:Substituents on the carbon atom at position 3, 17 and/or 19 includelower alkyl, for example, C₁₋₄ alkyl, especially methyl and ethyl.Substituents on the carbon atom at position 16 include lower alkyl e.g.methyl, ethyl etc., hydroxy and halogen atom e.g. chlorine, fluorine,aryloxy e.g. trifluoromethylphenoxy, etc. Substituents on the carbonatom at position 17 include halogen atom e.g. chlorine, fluorine etc.Substituents on the carbon atom at position 20 include saturated andunsaturated lower alkyl e.g. C₁₋₆ alkyl, lower alkoxy e.g. C₁₋₄ alkoxyand lower alkoxy (lower) alkyl e.g. C₁₋₄ alkoxy-C₁₋₄ alkyl. Substituentson the carbon atom at position 5 include halogen atom e.g. chlorine,fluorine etc. Substituents on the carbon atom at position 6 include oxogroup forming carbonyl. Stereochemistry of PGs having hydroxy, loweralkyl or lower (hydroxy) alkyl substituent on the carbon atom atposition 9 and/or 11 may be alpha, beta or mixtures thereof.

Especially preferred compounds are those having a lower alkyl e.g.methyl, ethyl, propyl, isopropyl, butyl, hexyl, preferably C₂₋₄ alkyland most preferably ethyl at position 20.

A group of preferred compounds used in the present invention has theformula ##STR2## wherein X and Y are hydrogen, hydroxy, halo, loweralkyl, hydroxy(lower)alkyl, or oxo, with the proviso that at least oneof X and Y is a group other than hydrogen, and 5-membered ring may haveat least one double bond, A is --COOH or its pharmaceutically acceptablesalt or ester, R₁ is bivalent saturated or unsaturated, lower or mediumaliphatic hydrocarbon residue which is unsubstituted or substituted withhalo, oxo or aryl, R₂ is saturated or unsaturated, medium aliphatichydrocarbon residue having 5 or more carbon atoms in the main orstraight chain moiety which is unsubstituted or substituted with halo,hydroxy, oxo, lower alkoxy, lower alkanoyloxy, cyclo(lower)alkyl, arylor aryloxy.

In the above formula, the term "unsaturated" in the definitions for R₁and R₂ is intended to include at least one and optionally more than onedouble bond and/or triple bond isolatedly, separately or seriallypresent between carbon atoms of the main and/or side chains. Accordingto usual nomenclature, an unsaturation between two serial positions isrepresented by denoting the lower number of said two positions, and anunsaturation between two distal positions is represented by denotingboth of the positions. Preferred unsaturation is a double bond atposition 2 and a double or triple bond at position 5.

The term "lower or medium aliphatic hydrocarbon residue" or "mediumaliphatic hydrocarbon residue" refers to a straight or branched chainhydrocarbyl group having 1 to 14 carbon atoms or 5 to 14 carbon atoms,respectively, (for a side chain, 1 to 3 carbon atoms being preferred)and preferably 2 to 8 carbon atoms for R₁ and 6 to 9 carbon atoms forR₂.

The term "halo" denotes fluoro, chloro, bromo and iodo.

The term "lower" throughout the specification is intended to include agroup having 1 to 6 carbon atoms unless otherwise specified.

The term "lower alkyl" as a group or a moiety in hydroxy(lower)alkyl,monocyclic aryl(lower) alkyl, monocyclic aroyl(lower)alkyl orhalo(lower)alkyl includes saturated and straight or branched chainhydrocarbon radicals containing 1 to 6, carbon atoms, e.g. methyl,ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl and hexyl.

The term "lower alkoxy" refers to the group lower-alkyl-O- wherein loweralkyl is as defined above.

The term "hydroxy(lower)alkyl" refers to lower alkyl as defined abovewhich is substituted with at least one hydroxy group, e.g.hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl and1-methyl-1-hydroxyethyl.

The term "lower alkanoyloxy" refers to a group of the formula: RCO--O--wherein RCO-- is an acyl group formed by oxidation of a lower alkylgroup as defined above, e.g. acetyl.

The term "cyclo(lower)alkyl" refers to a cyclic group formed bycyclization of a lower alkyl group as defined above.

The term "aryl" includes unsubstituted or substituted aromaticcarbocyclic or heterocyclic (preferably monocyclic) groups, e.g. phenyl,tolyl, xylyl and thienyl. Examples of substituents are halo andhalo(lower)alkyl wherein halo and lower alkyl being as defined above.

The term "aryloxy" refers to a group of the formula: ArO- wherein Ar isaryl as defined above.

Suitable "pharmaceutically acceptable salts" includes conventionalnon-toxic salts, and may be a salt with an inorganic base, for examplean alkali metal salt (e.g. sodium salt, potassium salt, etc.) and analkaline earth metal salt (e.g. calcium salt, magnesium salt, etc.),ammonium salt, a salt with an organic base, for example, an amine salt(e.g. methylamine salt, dimethylamine salt, cyclohexylamine salt,benzylamine salt, piperidine salt, ethylenediamine salt, ethanolaminesalt, diethanolamine salt, triethanolamine salt,tris(hydroxymethylamino)ethane salt, monomethyl-monoethanolamine salt,procaine salt, caffeine salt, etc.), a basic amino acid salt (e.g.arginine salt, lysine salt, etc.), tetraalkyl ammonium salt and thelike. These salts can be prepared by the conventional process, forexample from the corresponding acid and base or by salt interchange.

Examples of the "pharmaceutically acceptable esters" are aliphaticesters, for example, lower alkyl ester e.g. methyl ester, ethyl ester,propyl ester, isopropyl ester, butyl ester, isobutyl ester, t-butylester, pentyl ester, 1-cyclopropylethyl ester, etc., lower alkenyl estere.g. vinyl ester, allyl ester, etc., lower alkynyl ester e.g. ethynylester, propynyl ester, etc., hydroxy(lower) alkyl ester e.g.hydroxyethyl ester, lower alkoxy(lower)-alkyl ester e.g. methoxymethylester, 1-methoxyethyl ester, etc., and aromatic esters, for example,optionally substituted aryl ester e.g. phenyl ester, tosyl ester,t-butylphenyl ester, salicyl ester, 3,4-di-methoxyphenyl ester,benzamidophenyl ester etc., aryl(lower)alkyl ester e.g. benzyl ester,trityl ester, benzhydryl ester, etc. Examples of the amides are mono- ordi- lower alkyl amides e.g. methylamide, ethylamide, dimethylamide,etc., arylamide e.g. anilide, toluidide, and lower alkyl- oraryl-sulfonylamide e.g. methylsulfonylamide, ethylsulfonylamide,tolylsulfonylamide etc.

The term "pharmaceutically" is intended to be "ophthalmically" when usedin connection with an ophthalmic composition.

Preferred examples of A include --COOH, --COOCH₃, --COOCH₂ CH₃ and--COOCH(CH₃)₂.

The configuration of the ring and the α- and/or omega chain in the aboveformula (I) may be the same as or different from that in the primaryPGs. However, the present invention also includes a mixture of acompound having a primary configuration and that of an unprimaryconfiguration.

Examples of the typical compounds of the present invention are15-keto-PGs, 13,14-dihydro-15-keto-PGAs to Fs and their derivatives e.g.20-loweralkyl-derivatives, Δ² -derivatives, 3R,S-methyl-derivatives,6-xo-derivatives, 5R,S-fluoro-derivatives, 5,5-difluoro-derivatives,16R,S-methyl-derivatives, 16,16-dimethyl-derivatives,16R,S-fluoro-derivatives, 16,16-difluoro-derivatives,17S-methyl-derivatives, 17R,S-fluoro-derivatives,17,17-difluoro-derivatives and 19-methyl-derivatives.

The component (a) may be in the keto-hemiacetal equilibrium by forming ahemiacetal between hydroxy group at position 11 and ketone at position15.

The proportion of both tautomeric isomers, when present, variesdepending on the structure of the rest of the molecule or kind of anysubstituent present and, sometimes, one isomer may predominantly bepresent in comparison with the other. However, in this invention, it isto be appreciated that the compounds used in the invention include bothisomers. Further, while the compounds used in the invention may berepresented by a structure or name based on keto-form regardless of thepresence or absence of the isomers, it is to be noted that suchstructure or name does not intend elimination of the hemiacetal type ofcompounds.

In the present invention, any of the individual tautomeric isomers, amixture thereof, or optical isomers, a mixture thereof, a racemicmixture, and other isomers such as steric isomers can be used in thesame purpose.

Some of the compounds used in the present invention may be prepared bythe method disclosed in Japanese Patent Publications (unexamined) No.A-52753/1989, A-104040/1989, A-151519/1989 and A-108/1990.

Alternatively, these compounds may be prepared by a process analogous tothat described in the above publications in combination with the knownsynthetic method for the five-membered ring moiety.

In the process for preparing 13,14-dihydro-15-keto-compound:

A commercially available (-)-Corey lactone, which is used as a startingmaterial, is subjected to Collins oxidation to give an aldehyde. Thealdehyde is allowed to react with dimethyl (2-oxoalkyl)phosphonate anionto give an α,β-unsaturated ketone, and the resultant is reduced toketone. The carbonyl group of the ketone is allowed to react with a diolto give a ketal, thereby protected, then a corresponding alcohol isobtained by elimination of the phenylbenzoyl group, and the resultinghydroxy group is protected with dihydropyran to give a tetrapyranylether. Thus, precursors of PGs wherein the ω-chain is13,14-dihydro-15-keto-alkyl can be obtained.

Using the above tetrapyranyl ether as a starting material, 6-keto-PG₁ sof the formula: ##STR3## may be obtained as follows:

The tetrapyranyl ether is reduced using diisobutyl aluminium hydride andthe like to give a lactol, which is allowed to react with a ylideobtained from (4carboxybutyl)triphenylphosphonium bromide, and theresultant is subjected to esterification followed by cyclization,combining the 5,6-double bond and the C-9 hydroxyl group with NBS oriodine, providing a halide. The resultant is subjected todehydrohalogenation with DBU and the like to give a 6-keto compound,which is subjected to Jones oxidation followed by deprotection to givethe objective compound.

Further, PG₂ s of the formula: ##STR4## may be obtained as follows:

The above tetrapyranyl ether is reduced to the lactol, which is allowedto react with a ylide obtained from (4-carboxybutyl)triphenylphosphoniumbromide to give a carboxylic acid. The resultant is subjected toesterification followed by Jones oxidation and deprotection to give theobjective compound.

In order to obtain PG₁ s of the formula: ##STR5## using the abovetetrapyranyl ether as a starting material, in the same manner as PG₂ ofthe formula: ##STR6## the 5,6-double bond of the resulting compound issubjected to catalytic reduction followed by deprotection. To prepare5,6-dehydro-PG₂ s containing a hydrocarbon chain of the formula:##STR7## a monoalkyl copper complex or a dialkyl copper complex of theformula: ##STR8## is subjected to 1,4-addition with4R-t-butyldimethylsilyloxy-2-cyclopenten-1-one, and the resulting copperenolate is seized with 6-carboalkoxy-1-iodo-2-hexyne or a derivativethereof.

PGs containing a methyl group instead of a hydroxy group at the C-11position may be obtained as follows: PGA obtained by Jones oxidation ofthe hydroxy group at the C-9 position of the 11-tosylate is allowed toreact with a dimethyl copper complex to give 11-dehydroxy-11-methyl-PGE.Alternatively, an alcohol obtained after elimination of p-phenylbenzoylgroup is converted to a tosylate. An unsaturated lactone obtained by DBUtreatment of the tosylate is converted to a lactol. After introductionof an α-chain using Wittig reaction, the resulting alcohol (C-9position) is oxidized to give PGA. PGA is allowed to react with dimethylcopper complex to give 11-dehydroxy-11-methyl-PGE. The resultant isreduced using sodium borohydride and the like to give11-dehydroxy-11-methyl-PGF.

PGs containing a hydroxymethyl group instead of a hydroxyl group at theC-11 position is obtained as follow: 11-dehydroxy-11-hydroxymethyl-PGEis obtained by a benzophenone-sensitized photoaddition of methanol toPGA. The resultant is, for example, reduced using sodium borohydride togive 11-dehydroxy-11-hydroxymethyl-PGF.

16-Fluoro-PGs may be obtained using dimethyl(3-fluoro-2-oxoalkyl)phosphonate anion in the preparation of an α,β-unsaturated ketone. Similarly, 19-methyl-PGs may be obtained using adimethyl (6-methyl-2-oxoalkyl)phosphonate anion.

The preparations in the present invention are not construed to belimited to them, and suitable means for protection, oxidation, reductionand the like may be employed.

The β-adrenergic blockers used as the component (b) in the presentinvention refer to agents capable of blocking the β-adrenergic receptor.Typical examples of such agents are relatively less selectiveβ-adrenergic receptor blocking agents which are represented by thefollowing formula:

    A--OCH.sub.2 CH(OH)CH.sub.2 NHC(CH.sub.3)(R)

wherein A is an aromatic group and R is hydrogen atom or methyl.

The above group A includes 4-morpholino-1,2,5-thiadiazol-3-yl,2-acetylbenzofuran-7-yl, 1,2,3,4-tetrahydro-2-oxo-quinoline-5-yl.Preferred compounds include Timolol, Befunolol, Betaxolol, Levabunolol,Carteolol and pharmaceutically acceptable salts thereof such asinorganic salts, e.g. hydrochloride or organic salts, e.g. maleate.

Since the component (a) has an activity of lowering ocular pressurewithout accompanying transient ocular hypertension as shown by theprimary PGs, the combination of (a) and (b) can be used for thetreatment of various disease and conditions in which lowering of ocularpressure is desirous, for example glaucoma, ocular hypertension andother disease which accompanies increase in ocular pressure.

As used herein, the term "treatment" or "treating" refers to any meansof control of a disease in a mammal, including preventing the disease,curing the disease, relieving the disease and arresting or relieving thedevelopment of the disease.

The combination has an advantage, by containing the component (b) inaddition to the component (a), that it has a synergistically increasedocular hypotensive action, thus enabling reduce in dosage, and/orlowering the side-effect.

The ratio (a):(b) in the combination varies, without limitation,ordinarily within the range 1:0.5 to 1:200, preferably 1:1 to 1:100 andmost preferably 1:2 to 1:50.

While the dosage of the component (a) varies depending on condition ofthe component(a) varies depending on condition of the patient, severityof the disease, purpose of the treatment, judgement of the physician andtotal dosage of the combination, it is ordinarily within the range 0.005to 2% and preferably 0.01 to 1% by weight.

The dosage of the component (b) varies, for example, depending on theconcentration of the component (a) and ordinarily within the range 0.005to 20% and preferably 0.01 to 10% by weight.

The combination according to the present invention can be administeredin the form of a pharmaceutical composition containing the components(a) and (b) and optionally other ingredients conveniently used in theophthalmic composition, such as carrier, diluent or excipient.

The ophthalmic composition used according to the invention includesliquids such as ophthalmic solution, emulsion, dispersion etc. andsemisolids such as ophthalmic gel, ointment etc. Diluents for theaqueous solution or suspension include, for example, distilled water andphysiological saline. Diluents for the nonaqueous solution andsuspension include, for example, vegetable oils e.g. olive oil, liquidparaggine, mineral oil, and propylene glycol and p-octyldodecanol. Thecomposition may also contain isotonization agents such as sodiumchloride, boric acid, sodium citrate, etc. to make isotonic with thelacrimal fluid and buffering agents such as borate buffer, phosphatebuffer, etc. to maintain pH about 5.0 to 8.0. Further, stabilizers suchas sodium sulfite, propylene glycol, etc., chelating agents such assodium edetate, etc., thickeners such as glycerol,carboxymethylcellulose, carboxyvinyl polymer, etc. and preservativessuch as methyl paraben, propyl paraben, etc. may also be added. thesecan be sterilized e.g. by passing through a bacterial filter or byheating.

The ophthalmic ointment may contain vaseline, Plastibase, Macrogol, etc.as a base and surfactant for increasing hydrophilicity. It may alsocontain geling agents such as carboxymethylcellulose, methylcellulose,carboxyvinyl polymer, etc.

In addition, the composition may contain antibiotics such aschloramphenicol, penicilin, etc. in order to prevent or treat bacterialinfection.

A more complete understanding of the present invention can be obtainedby reference to the following Preparation Examples, Formulation Examplesand Test Examples which are provided herein for purpose of illustrationonly and are not intended to limit the scope of the invention.

Preparations

Preparations of 13,14-dihydro-15-keto-20-ethyl-PGA₂ isopropyl ester,13,14-dihydro-15-keto-20-ethyl-PGE₂ isopropyl ester and13,14-dihydro-15-keto-20-ethyl-PGF₂ α isopropyl ester (cf. Preparationchart I):

1) Preparation of 1S-2-oxa-3-oxo-6R-(3-oxo-1-transdecenyl)-7R-(4-phenylbenzoyloxy)-cis-bicyclo[3.3.0]-octane (3):

Commercially available (-)-Corey lactone (1) (7 g) was subjected toCollins oxidation in dichloromethane to give aldehyde (2). The resultantwas allowed to react with dimethyl (2-oxononyl)phosphonate (4.97 g)anion to give1S-2-oxa-3-oxo-6R-(3,3-ethylendioxy-1-trans-decenyl)-7R-(4-phenylbenzoyloxy)-cis-bicyclo[3.3.0]-octane(3).

2) Preparation of1S-2-oxa-3-oxo-6R-(3-oxodecyl)-7R-(4-phenylbenzoyloxy)-cis-bicyclo[3.3.0]-octane(4):

Unsaturated ketone (3) (7.80 g) was reduced in ethyl acetate (170 ml)using 5% Pd/C under hydrogen atmosphere. The product obtained after theusual work-up (4) was used in the following reaction.

3) Preparation of1S-2-oxa-3-oxo-6R-(3,3-ethylenedioxy-decyl)-7R-(4-phenylbenzoyloxy)-cis-bicyclo[3.3.0]-octane(5):

Saturated ketone (4) was converted to ketal (5) in dry benzene (150 ml)using ethylene glycol and p-toluenesulfonic acid (catalytic amount).

4) Preparation of1S-2-oxa-3-oxo-6R-(3,3-ethylenedioxy-decyl)-7R-hydroxy-cis-bicyclo[3.3.0]-octane(6):

To a solution of ketal (5) in absolute methanol (150 ml) was addedpotassium carbonate (2.73 g). The mixture was stirred overnight at roomtemperature. After neutralization with acetic acid, the resultant wasconcentrated under reduced pressure. The resulting crude product wasextracted with ethyl acetate. The organic layer was washed with a diluteaqueous solution of sodium bicarbonate and a saline, and dried. Thecrude product obtained after evapolation was chromatographed to givealcohol (6). Yield; 3.31 g

5) Preparation of lactol (7):

Alcohol (6) (0.80 g) was reduced in dry toluene (8 ml) using DIBAL-H at-78° C. to give lactol (7). 6) Preparation of13,14-dihydro-15,15-ethylenedioxy-20-ethyl-PGF₂ α (8):

A DMSO solution of lactol (7) was added to ylide prepared from(4-carboxybutyl)triphenylphosphonium bromide (3.65 g). The reactionmixture was stirred overnight to give carboxylic acid (8).

7) Preparation of 13,14-dihydro-15,15-ethylenedioxy-20-ethyl-PGF₂ αisopropyl ester (9):

Carboxylic acid (8) was converted to13,14-dihydro-15,15-ethylenedioxy-20-ethyl-PGF₂ α isopropyl ester (9)using DBU and isopropyl iodide in acetonitrile.

Yield; 0.71 g

8) Preparation of 13,14-dihydro-15-keto-20-ethyl-PGF₂ α isopropyl ester(10):

13,14-Dihydro-15,15-ethylenedioxy-20-ethyl-PGF₂ α isopropyl ester (9)(0.71 g) was kept in acetic acid/THF/water (3/1/1) at 40° C. for 3hours. The crude product obtained after concentration under reducedpressure was chromatographed to give 13,14-dihydro-15-keto-20-ethyl-PGF₂α isopropyl ester (10).

Yield; 0.554 g

9) Preparation of 13,14-dihydro-15-keto-20-ethyl-PGA₂ isopropyl ester(12):

A solution of 13,14-dihydro-15-keto-20-ethyl-PGF₂ α isopropyl ester (10)(0.125 g) and p-toluenesulfonyl chloride (0.112 g) in pyridine (5 ml)was maintained at 0° C. for 2 days. According to the usual work-up,tosylate (11) was obtained.

Tosylate (11) was subjected to Jones oxidation in acetone (8 ml) at -25°C. The crude product obtained after the usual work-up waschromatographed to give 13,14-dihydro-15-keto-20-ethyl-PGA₂ α isopropylester (2).

Yield; 0.060 g

10) Preparation of13,14-dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGF.sub.2α isopropyl ester (13):

13,14-Dihydro-15,15-ethylenedioxy-20-ethyl-PGF₂ α isopropyl ester (9)(3.051 g) was dissolved in dry N,N-dimethylformamide (25 ml),t-butyldimethylsilyl chloride (1.088 g) and imidazole (0.49 g) was addedthereto. The resultant was stirred at room temperature overnight. Thereaction mixture was concentrated under reduced pressure, and theresulting crude product was chromatographed to give13,14-dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGF.sub.2α isopropyl ester (13).

Yield; 2.641 g

11) Preparation of13,14-dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGE.sub.2isopropyl ester (14):

13,14-Dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGF.sub.2α isopropyl ester (13) (1.257 g) was subjected to Jones oxidation at±40° C. After the usual work-up, the resulting crude product waschromatographed to give13,14-dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGE.sub.2isopropyl ester (14).

Yield; 1.082 g

12) Preparation of 13,14-dihydro-15-keto-20-ethyl-PGE₂ isopropyl ester(15):

To a solution of13,14-dihydro-15,15-ethylenedioxy-20-ethyl-11-t-butyldimethylsiloxy-PGE.sub.2isopropyl ester (14) in acetonitrile was added hydrofluoric acid (46%aqueous solution). The mixture was stirred at room temperature for 40minutes. The crude products obtained after usual work-up waschromatographed to give 13,14-dihydro-15-keto-20-ethyl-PGE₂ isopropylester (15).

Yield; 0.063 g (97%) ##STR9##

TEST EXAMPLE 1

Japanese white rabbits (weight: 2.5-3.5 kg, 6 animals/group) were fixedand eyes were anesthetized by dropping 0.4% oxybuprocaine hydrochlorideto eyes. The ocular pressure measured at 0.5-1 hour after the fixationwas taken as the 0 hour value and values of pressure thereafter weremeasured in the course of time administering by eye-dropping each 50 μlof the following formulations. An electronic pneumatonometer (Alcon) wasused for measurement. Decrease in ocular pressure (mean value) at 5hours after administration of each of the formulations was compared inthe Table 1.

Formulation Example 1 (Comparative)

    ______________________________________                                        Timolol maleate     0.1       g                                               Sterilized water    q.s. to 100                                                                             ml                                              ______________________________________                                    

Formulation Example 2 (Comparative)

    ______________________________________                                        Isopropyl (Z)-7-[(1R,2R,3R,5S)-3,5-                                                                  0.01      g                                            dihydroxy-2-(3-oxodecyl)cyclopentyl]-                                         hept-5-noate [13,14-dihydro-15-keto-                                          20-ethyl-PGF.sub.2 α isopropyl ester,                                   hereinafter referred to as Compound A]                                        Sterilized water       q.s. to 100                                                                             ml                                           ______________________________________                                    

Formulation Example 3

    ______________________________________                                        Timolol maleate     0.1       g                                               Compound A          0.01      g                                               Sterilized water    q.s. to 100                                                                             ml                                              ______________________________________                                    

                  TABLE 1                                                         ______________________________________                                                  Decrease in ocular pressure (mmHg)                                  ______________________________________                                        Formulation 1                                                                             -0.2                                                              Formulation 2                                                                             -0.3                                                              Formulation 3                                                                             -3.1                                                              ______________________________________                                    

The above results show that the combined use of Timolol maleate andCompound A result in a synergistic effect.

What we claim is:
 1. A method for treatment of ocular hypertension whichcomprises ocularly administering, to a subject in need of suchtreatment, an oculo-hypotensively synergistic combination of(a) a13,14-dihydro-15-ketoprostaglandin compound, and (b) a β-adrenergicblockerin an amount effective in treatment of ocular hypertension. 2.The method according to claim 1, in which the component (a) is a13,14-dihydro-15-ketoprostaglandin A, B, C, D, E or F, or apharmaceutically acceptable salt thereof, or a pharmaceuticallyacceptable ester thereof.
 3. The method according to claim 1, in whichthe component (a) is a 13,14-dihydro-15-keto-20-loweralkylprostaglandin,or a pharmaceutically acceptable salt thereof, or a pharmaceuticallyacceptable ester thereof.
 4. The method according to claim 1, in whichthe component (a) is a 13,14-dihydro-15-keto-20-ethylprostaglandin, or apharmaceutically acceptable salt thereof, or a lower alkyl esterthereof.
 5. The method according to claim 1, in which the component (a)is a 13,14-dihydro-15-keto-20-loweralkylprostaglandin F₂ α or apharmaceutically acceptable salt thereof, or a lower alkyl esterthereof.
 6. The method according to claim 1, in which the component (a)is a 13,14-dihydro-15-keto-20-ethylprostaglandin F₂ α, or apharmaceutically acceptable salt thereof, or a lower alkyl esterthereof.
 7. The method according to claim 1, in which the component (b)is Timolol, Befunolol, Betaxolol, Levobunolol, Carteolol, or apharmaceutically acceptable salt thereof.
 8. The method according toclaim 1, in which the components (a) and (b) are administered in theratio (a):(b) of 1:0.5 to 1:200.
 9. The method according to claim 1, inwhich the components (a) and (b) are administered simultaneously orsequentially.
 10. A pharmaceutical composition for treatment of ocularhypertension comprising an oculo-hypotensively synergistic combinationof(a) a 13,14-dihydro-15-ketoprostaglandin compound, and (b) aβ-adrenergic blockerin association with a pharmaceutically acceptablecarrier, diluent or excipient.
 11. The method according to claim 3, inwhich the component (b) is represented by the formula:

    A--OCH.sub.2 CH(OH)CH.sub.2 NHC(CH.sub.3).sub.2 (R)

wherein A is an aromatic group and R is hydrogen atom or methyl.
 12. Themethod according to claim 4, n which the component (b) is represented bythe formula:

    A--OCH.sub.2 CH(OH)CH.sub.2 NHC(CH.sub.3).sub.2 (R)

wherein A is an aromatic group and R is hydrogen atom or methyl.
 13. Themethod according to claim 5, in which the component (b) is representedby the formula:

    A--OCH.sub.2 CH(OH)CH.sub.2 NHC(CH.sub.3).sub.2 (R)

wherein A is an aromatic group and R is hydrogen atom or methyl.
 14. Themethod according to claim 6, in which the component (b) is representedby the formula:

    A--OCH.sub.2 CH(OH)CH.sub.2 NHC(CH.sub.3).sub.2 (R)

wherein A is an aromatic group and R is hydrogen atom or methyl.
 15. Themethod according to claim 3, in which the component (b) is Timolol,Befunolol, Betaxolol, Levobunolol, Carteolol, or a pharmaceuticallyacceptable salt thereof.
 16. The method according to claim 4, in whichthe component (b) is Timolol, Befunolol, Betaxolol, Levobunolol,Carteolol, or a pharmaceutically acceptable salt thereof.
 17. The methodaccording o claim 5, in which the component (b) is Timolol, Befunolol,Betaxolol, Levobunolol, Carteolol, or a pharmaceutically acceptable saltthereof.
 18. The method according to claim 6, in which the component (b)is Timolol, Befunolol, Betaxolol, Levobunolol, Carteolol, or apharmaceutically acceptable salt thereof.
 19. The method according toclaim 3, in which the component (b) is Timolol, or a pharmaceuticallyacceptable salt thereof.
 20. The method according to claim 4, in whichthe component (b) is Timolol, or a pharmaceutically acceptable saltthereof.
 21. The method according to claim 5, in which the component (b)is Timolol, or a pharmaceutically acceptable salt thereof.
 22. Themethod according to claim 6, in which the component (b) is Timolol, or apharmaceutically acceptable salt thereof.